Gene Therapy Research Unit (GTRU)

Lab head: Dr Ian Alexander
Location: Children¿s Medical Research Institute, Westmead

The GTRU is a joint initiative between the Children’s Medical Research Institute and The Children’s Hospital at Westmead.  Headed by Professor Ian Alexander, a clinician/scientist with a strong research interest in genetic disease, the unit is focused on developing gene therapy strategies to treat disorders of the liver and bone marrow.

Lab members: Prof Ian E Alexander (Head)
Research approach equipment: Since its inception in 1995, the unit has systematically established the resources and infrastructure to make fundamental discoveries in basic science and translate the findings via preclinical models from laboratory bench to patient bedside. The research is always guided by a ¿ground-up¿ approach to realise the long-term vision to implement gene therapies in the clinic.

  1. Baoutina, A., Alexander, I.E., Rasko, J.E.J., and Emslie, K.R. (2007). Potential use of gene therapy in athletic performance enhancement. Molecular Therapy 15(10):1751-1766, epub 7th August, 2007.
  2. Baoutina, A., Alexander, I.E., Rasko, J.E.J., and Emslie, K.R. (2008). Developing strategies for detection of gene doping. Journal of Gene Medicine 10(1):3-20.
  3. Curtin J.A., Dane A.P., Swanson A., Alexander I.E. and Ginn S.L. (2008) Bi-directional promoter interference between two widely used internal heterologous promoters in a late-generation lentiviral construct. Gene Therapy 15(5):384–390, epub 24th January, 2008.
  4. Alexander I.E., Cunningham S.C., Logan G.L. and Christodoulou J. (2008) Potential of AAV Vectors in the Treatment of Metabolic Disease. Invited Review. Gene Therapy 15(11):831-839, epub 10th April, 2008.
  5. Cunningham S.C., Dane A.P., Spinoulas A., Logan G.J. and Alexander I.E. (2008) Gene delivery to the juvenile mouse liver using AAV2/8 vectors. Molecular Therapy 16(6):1081-1088, epub 15th April, 2008.
  6. Lee V.W., Wang Y-M., Wang Y-P, Zheng D., Polhill T., Cao Q., Wu H., Alexander I.E., Alexander S.I. and Harris D.C. (2008) Regulatory immune cells in kidney disease. American Journal of Physiology: Renal Physiology 295(2):F335-342, epub 16th April, 2008.
  7. Ginn S.L. and Alexander I.E. (2008) Gene Therapy. In: Wiley Encyclopedia of Clinical Trials. John Wiley & Sons, Inc., New Jersey, Eds. D’Agostino R., Sullivan L., Massaro J.
  8. Laurence J.M., Wang C., Cunningham S.C., Zheng M., Earl J., Tay S.S., Allen R.D.M., McCaughan G.W., Alexander I.E., Bishop G.A. and Sharland A.F. (2009) Over-expression of indoleamine dioxygenase in rat liver allografts using a high-efficiency adeno-associated virus vector does not prevent acute rejection. Liver Transplantation 15(2):233-241.
  9. Logan G.J., Wang L., Zheng M., Ginn S.L., Coppel R.L. and Alexander I.E. (2009) Antigen-specific humoral tolerance or immune augmentation induced by intramuscular delivery of adeno associated virus encoding CTLA4-Ig-antigen fusion molecules. Gene Therapy 16(2):200-210, epub 27th November, 2008.
  10. Ginn S.L., Cunningham S.C., Zheng M., Spinoulas A., Carpenter K.H. and Alexander I.E. (2009) In vivo assessment of mutations in OTC for dominant-negative effects following rAAV2/8-mediated gene delivery to the mouse liver. Gene Therapy 16(6): 820-823, epub 9th April, 2009.
  11. Laurence J.M., Allen R.D.M., McCaughan G.W., Logan G.J., Alexander I.E., Bishop G.A., Sharland A.F. (2009) Gene Therapy in Transplantation. Transplantation Reviews 23(3):159-170,epub 8th May, 2009.
  12. Cunningham S.C., Spinoulas A., Carpenter K., Wilcken B., Kuchel P. and Alexander I.E. (2009) AAV2/8-mediated correction of OTC deficiency is robust in adult but not neonatal spfash mice. Molecular Therapy, 17(8):1340-1346, epub 21st April 2009.
  13. Dane A.P., Cunningham S.C., Graf N.S. and Alexander I.E. (2009) Sexually Dimorphic Patterns of Episomal rAAV Genome Persistence in the Adult Mouse Liver and Correlation with Hepatocellular Proliferation. Molecular Therapy, 17(9):1548-1554, epub 30th June, 2009.
  14. Deakin C., Alexander I.E. and Kerridge I. (2009) Accepting Risk in Clinical Research: Is Gene Therapy Becoming Too Risk-Averse? Molecular Therapy, 17(11):1842-1848, epub 22nd September, 2009.
  15. Logan G.J., Wang L., Zheng M., Coppel R.L. and Alexander I.E. (2010) Antigen fusion with C3d3 augments or inhibits humoral immunity to AAV genetic vaccines in a transgene-dependent manner. Immunology and Cell Biology, 88(2):228-232, epub 24th November, 2009.
  16. Ginn S.L, Liao S.H.Y., Dane A.P., Smyth C., Hyman J., Finnie J.W., Zheng M.Z., Thrasher A.J. and Alexander I.E. (2010) Lymphomagenesis in SCID-X1 mice following lentivirus mediated phenotype correction independent of insertional mutagenesis and gc overexpression. Molecular Therapy, 18(5):965-976, epub 30th March, 2010. Subject of editorial comment.
  17. Sharland A., Logan G.J., Bishop G.A. and Alexander I.E. (2010) Liver-directed Gene Expression Using Recombinant AAV 2/8 Vectors - a Tolerogenic Strategy for Gene Delivery? Discovery Medicine, 9(49):519-527, epub 9th June, 2010.
  18. Deakin C., Alexander I.E. and Kerridge I. (2010) The Ethics of Gene Therapy: Balancing the Risks. Current Opinion in Molecular Therapeutics 12(5):578-585.
  19. Cunningham S.C., Kok C.Y., Dane A.P., Carpenter K., Kizana E., Kuchel P.W. and Alexander I.E. (2011) Induction and prevention of severe hyperammonaemia in the spfash mouse model of ornithine transcarbamylase deficiency using shRNA and rAAV-mediated gene delivery. Molecular Therapy, 19(5):854-859, epub 8th March 2011.
  20. Chung H., Lin R.C.Y., Logan G.J., Alexander I.E., Sachdev P.S. and Sidhu K.S. (2012) Human induced pluripotent stem cells derived under feeder-free conditions display unique cell cycle and DNA replication gene profiles. Stem Cells and Development, 21(2):206-216, epub 1st June 2011.
  21. Alexander I.E., Kok C., Dane A.P. and Cunningham S.C. (2012) Gene Therapy for Metabolic Disorders:  An overview with a focus on urea cycle disorders.  J. Inher. Meta. Dis., 35(4):641-645, epub 9th March 2012.
  22. Ginn S.L. and Alexander I.E. (2012) Gene therapy: Progress in childhood disease.  Invited review - Journal of Paediatrics and Child Health, 48(6):466-471, epub 21st Oct 2011.
  23. Logan, G.J. and Alexander I.E. (2012) Adeno-Associated Virus vectors: Immumobiology and potential use for immune modulation. Current Gene Therapy, 12(4):333-343.
  24. Dane A.P., Wowro S.J., Cunningham S.C. and Alexander I.E. (2013) Comparison of gene transfer to the murine liver following intraperitoneal and intraportal delivery of hepatotropic AAV pseudo-serotypes. Gene Therapy, 20(4):460-464, epub 16th August 2012.
  25. Cunningham E.C., Tay S.S., Wang C., Rtshiladze M., Wang Z.Z., McGuffog C., Cubitt J.,McCaughan G.W., Alexander I.E., Bowen D., Sharland A.F., Bertolino P.D. and Bishop G.A. (2013) Gene Therapy for tolerance: High-level expression of donor MHC in the liver overcomes naïve and memory alloresponses to skin grafts. Transplantation 95(1):70-77, epub December 2012.
  26. Ginn S.L., Alexander I.E., Edelstein M.L., Abedi M.R. and Wixon J. (2013) Gene therapy clinical trials worldwide to 2012 - an update. Invited Review - Journal of Gene Medicine 15: 65-77, epub 29th January 2013.
  27. Deakin C.T., Alexander I.E., Hooker C.A. and Kerridge I.H. (2013)  Gene therapy researchers’ assessments and perceptions of risks in clinical trials.  Molecular Therapy 21(4): 806-815,epub 22nd January 2013.
  28. Gerace D., Ren B., Hawthorne W.J., Byrne M.R., Phillips P., O’Brien B.A., Nassif N., Alexander I.E. and Simpson A.M. (2013) Pancreatic Transdifferentiation in Porcine Liver Following Lentiviral Delivery of Human Furin-Cleavable Insulin. Transplantation Proceedings 45(5): 1869-1874.
  29. Ginn S.L. and Alexander I.E. (2013) Gene Therapy. In: Methods and Applications of Statistics in Clinical Trials, Volume 1: Concepts, Principles, Trials and Design (In press).
  30. Kok C.Y., Cunningham S.C., Carpenter K.H., Dane A.P., Siew S.M., Logan G.J., Kuchel P.W. and Alexander I.E. (2013) Adeno associated virus-mediated rescue of neonatal lethality in argininosuccinate synthetase deficient mice. Molecular Therapy , epub 2nd July 2013.

Repairing genetic mutations in the liver stem cell using recombinant adeno-associated virus (AAV)

Primary supervisor: Ian Alexander

Gene therapy is beginning to fulfil its potential in the clinic for the treatment of monogenic disorders.  All reports to date rely on a “gene addition” approach such that diseased cells are modified to encode the correct copy of an affected gene.  The ideal strategy , however, is “gene repair” so that gene expression is maintained under endogenous regulatory elements.  In this respect, AAV-mediated homologous recombination (HR) demonstrates 1000-fold greater efficiencies for gene repair over alternative approaches that use plasmid DNA and adenovirus.  Despite the greater efficiencies mediated by AAV gene repair, the system requires further improvement to provide therapeutic utility.

The aim of this project is to optimise AAV mediated homologous recombination to repair mutant loci in liver stem cells (LPC), a cell type with therapeutic potential to treat metabolic liver disease.  LPC can be propagated and expanded ex vivo whilst retaining the ability to terminally differentiate into hepatocytes for engraftment in the diseased liver.   Our laboratory has made considerable progress with this project which offers experience in a range of laboratory techniques including molecular biology, packaging of recombinant viruses, tissue culture, histology and animal studies.

Discipline: Pathology
Co-supervisors: Grant Logan
Keywords: Gene therapy, Stem cells, Liver